The pattern of distribution in rat spinal cord and changing pattern during normal aging of c-Fos, Bcl-2, Bax, and p53 expression were investigated by immunohistochemical staining. Male Sprague-Dawley rats at the age of one week, five months, and two years were studied. C-Fos immunoreactivity was observed diffusely in gray matters in neonatal rats, preferentially located in deep dorsal horn and around central canal. Compared with those of neonatal rats, immunoreactive cells decreased prominently in adult rats. In aged rats, these cells were not seen in any segments. In a transverse section, spatial expression of Bcl-2 and Bax proteins showed a diffuse distribution pattern with immunore-activity more prominent in the anterior horn. Continuing expression of these proteins was shown in each age group. In adult rats, Bcl-2 immunoreactivity was decreased drastically compared to that of neonatal rats. The immunoreactivity was higher in aged than in adult rats, but the number of immunoreactive cells was not different between aged and neonatal rats. The number of Bax-immunoreactive cells was greater in adult than in neonatal rats; in aged rats, it was similar with that of adult rats. The spinal cords of neonatal rats were not p53-immunoreactive, though p53-positive cells were detected in all segments of adult spinal cord. P53-positive cells were stained along the cellular margin, with a pale central portion. The pattern of p53 immunoreactivity in adult and aged rats was similar; the number of p53-positive cells, however, was higher in aged rats than in adult. In conclusion, we demonstrated that the expression patterns of c-Fos, Bcl-2, Bax, and p53 proteins in rat spinal cord change during normal aging for the first time.
Adult ; Aging* ; Animals ; bcl-2-Associated X Protein ; Horns ; Humans ; Male ; Rats* ; Rats, Sprague-Dawley ; Spinal Cord*

Nitric oxide (NO) involvement has been demonstrated in mechanisms of synaptic plasticity, particularly in hippocampal long-term potentiation, a mechanism that underlies certain forms of learning and memory. Further, NO has been shown to regulate various neurotransmitters which play an important role in learning and memory. Several findings suggest that NO production may be decreased in the aged rat. Changes in the nNOS-containing neurons with aging were demonstrated by immunocytochemistry and in situ hybridization. NOS-immunoreactive cells in aged rats were present in all cortical areas and the hippocampus, and the pattern of distribution was similar to that of the control group. The number of NOS-immunoreactive cells in the cerebral cortex was significantly decreased in the aged rats, but the extent of changes was variable in each area, and ranged from mild decrease (<30%) to severe decrease (>50%). Severely decreased areas were the cingulate cortex, parietal cortex area 1, temporal cortex area 1, 2, 3, medial part of occipital cortex area 2, monocular and binocular part of occipital cortex area 1, entorhinal cortex, hippocampus proper, dentate gyrus and subiculum. Moderately decreased areas (30~50%) were frontal cortex area 1, 2, 3, parietal cortex area 2, forelimb, hindlimb, lateral part of occipital cortex area 2. Slightly decreased area was insular cortex. Morphologically, the number of dendritic branches seemed to be decreased in aged group and the length of dendrites of NOS-IR neurons showed a tendency to shorten. These results indicate the involvement of neuronal system containing NOS in the aging brain, and provide the first morphological evidence for the loss of NOS neurons in the cerebral cortex of the aged rats by immunocytochemistry. Further multidisciplinary investigations involving normal aging and neurodegenerative disease such as Alzheimer's disease are needed to clarify the importance of nitric oxide changes in the cerebral cortex with aging.
Aging ; Alzheimer Disease ; Animals ; Brain ; Cerebral Cortex* ; Dendrites ; Dentate Gyrus ; Entorhinal Cortex ; Forelimb ; Gyrus Cinguli ; Hindlimb ; Hippocampus ; Immunohistochemistry ; In Situ Hybridization ; Learning ; Long-Term Potentiation ; Memory ; Neurodegenerative Diseases ; Neurons* ; Neurotransmitter Agents ; Nitric Oxide ; Plastics ; Rabeprazole ; Rats* ; Telescopes

Long-Evans Cinnamon (LEC) mutant rat, which spontaneously develops a necrotizing hepatic injury at 4 ~5 months of age, reveals an excess hepatic copper accumulation and is a good model for studying the detail mechanism of cellular copper toxicity. We have observed the effects of copper toxicity on DNA synthesis upon growth stimulation by treating primary-cultured hepatocytes of LEC rat with epidermal growth factor (EGF) and insulin. DNA synthesis measured by [ 3 H]-thymidine incorporation and DNA synthesis S-phase cells in LEC rat significantly decreased when compared to those of normal F344 rat. Since DNA synthesis was impaired in LEC rat, we examined the detail mechanism by determining the histone content, which are involved in DNA stability, and the phosphorylation of nuclear protein. However, the histone contents and phosphorylated nuclear protein upon growth stimulation was intact in LEC rat. These results suggest that a cellular event other than protein phosphorylation required for the initiation of DNA synthesis upon growth stimulation is impaired by copper cytotoxicity in LEC rat.
Animals ; Cinnamomum zeylanicum ; Copper* ; DNA* ; Epidermal Growth Factor ; Hepatocytes ; Histones ; Insulin ; Nuclear Proteins ; Phosphorylation ; Rats* ; Rats, Inbred F344

Maintenance of cellular iron homeostasis is a prerequisite for proliferation and differentiation of cells, and is also a central role in the regulation of immune function. Monocyte-macrophages play an important roles in host defense, particularly in the inflammatory process of acute and chronic disease. The reason that an iron is important in these cell is because an iron is indispensable in a generation of hydroxyl radical for bacterium killing. Because of the role of iron in the monocytic THP-1 cell differentiation is not become clear, we investigated whether THP-1 cell can differentiate to macrophage-like cell using of iron and iron chelator which cause iron depletion. The cell differentiation was not able to observe by iron treatment, by the way, the cell adhesion was increased in DFO treated monocyte and cellular pseodopodial extension, change of a nucleus-cytoplasmic ratio were showed in Differential interference contrast (DIC) and Giemsa staining, and it was inhibited by ferric citrate (FC). Increased polystyrene bead phagocytosis by DFO treatment of THP-1 cell were detected through FACS and rhodamine-phallodin staining. The SR-A expression, which was a cell differentiation marker, was increased by DFO treatment of THP-1 cell. These results suggest that iron depletion by DFO can promote THP-1 cell diffentiation into macrophage-like cell, and this may carrying out important role in the immune response.
Azure Stains ; Cell Adhesion ; Cell Differentiation ; Chronic Disease ; Citric Acid ; Deferoxamine ; Homeostasis ; Homicide ; Hydroxyl Radical ; Iron* ; Macrophages ; Monocytes ; Phagocytosis ; Polystyrenes

Chromosomal abnormalities, which are valuable markers for diagnosis and prognosis of cancer, provide useful clues in characterizing cancer at molecular level. Gastric cancer is the major cause of cancer deaths in Asian countries, including Korea. Genetic changes during the progression and metastasis of gastric cancer remain unclear. Recently, technique of degenerate oligonucleotide primed (DOP) PCR-comparative genomic hybridization (CGH) permits genetic imbalances screening of the entire genome using only small amounts of tumor DNA. In non-metastatic gastric cancers the common sites of copy number increases were detected at 8q (64%), 4p12-q24 (64%), 5p13-q23 (64%), 13q21-q32 (64%), 6q11-q21 (55%), 7q(50%), 14q11.2-q21 (45%), 3q11-q13.3 (41%), and 2q23-q32 (41%). In metastatic gastric cancers, the frequent sites of gains were detected at 8p21-qter (60%), 5 (54%), 20 (42%), 6pter-q24 (51%), 1q21-qter (46%), 3p14-qter (46%), 22q (46%), and 4 (43%). Deletion or chromosomal loss was found to be less frequent in this study. The frequent sites of copy number decreases were detected at 1p34-pter (23%), 16q23-q24 (18%), and 19q13 (18%) in non-metastatic gastric cancers. In metastatic gastric cancers, chromosome losses were detected at X (37%), 1p33-pter (37%), and 16p (23%). The recurrent gains and losses of chromosomal regions identified in this study provide candidate regions that may contain oncogenes or tumor suppressor genes respectively involved in the tumorigenesis of gastric cancer.
Asian Continental Ancestry Group ; Carcinogenesis ; Chromosome Aberrations ; Comparative Genomic Hybridization* ; Diagnosis ; DNA ; Genes, Tumor Suppressor ; Genome ; Humans ; Korea ; Mass Screening ; Neoplasm Metastasis ; Nucleic Acid Hybridization ; Oncogenes ; Prognosis ; Stomach Neoplasms

Nerve regeneration in the central nervous system has been studied by grafting various tissues and cells. Choroid plexus epithelial cells represent a continuation of ventricular ependymal cells and have the same origin as regarded as modified ependymal cells. To study the use of choroid plexus ependymal cell grafting for nerve regeneration in the spinal cord, the choroid plexus was excised from the lateral and fourth ventricles of adult Sprague-Dawley rats, minced into small fragments, and grafted at the T9 level in adult rat spinal cord transected or contused. In this study, transplants of choroid plexus ependymal cells were successfully used to promote functional and structural recovery after spinal cord transection and contusion. The area of damaged spinal cord was diminished after choroid plexus ependymal cells transplantation. Nearly normal anterior horn cells were observed immediately distal to the transected region. Tyrosine hydroxylase immunoreactive descending fibers were observed in the distal region beyond transected area. These findings indicate that choroid plexus ependymal cells have the ability to facilitate axonal growth, suggesting that they may be a promising candidate as graft for the promotion of nerve regeneration in the spinal cord.
Adult ; Animals ; Anterior Horn Cells ; Axons ; Cell Transplantation* ; Central Nervous System ; Choroid Plexus ; Contusions ; Epithelial Cells ; Fourth Ventricle ; Humans ; Nerve Regeneration* ; Rats* ; Rats, Sprague-Dawley ; Spinal Cord Injuries* ; Spinal Cord* ; Transplants* ; Tyrosine 3-Monooxygenase

Heme oxygenase-1 (HO-1) is a stress-inducible enzyme with anti-inflammatory activity, but the mechanisms underlying this activity are incompletely understood. Nuclear transcription factor kappa B (NF-kappa B) activation is an important factor in the pathogenesis of inflammatory bowel disease (IBD). We investigated the suppressive effects of HO-1 on the activation of NF-kappa B by pro-inflammatory cytokines in cultured colonic epithelial cells and by trinitrobenzene sulfonic acid (TNBS) in the colon of mice. The expression level of HO-1 in the colonic epithelium of a patient with inflammatory bowel disease and pseudo-membranous colitis was lower than that in a healthy control subject. In cultured human colonic epithelial HT-29 cells, pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha ) and IL-1 beta down-regulate HO-1 expression. The HO-1 inducer, cobalt protoporphyrin IX (CoPPIX), dramatically down-regulated NF-kappa B activation in HT-29 cells by TNF-alpha. In addition, bilirubin-a product of heme catabolism by HO-1-and the carbon monoxide donor tricarbonyldichlororuthenium (II) dimer also suppressed NF-kappa B activation by TNF-alpha. However, iron, another heme metabolite, did not suppress NF-kappa B activation by TNF-alpha. Furthermore, CoPPIX diminished the macroscopic and histopathological symptoms of TNBS-induced colitis and down-regulated NF-kappa B activation in mice. In conclusion, this study suggests that HO-1 plays an important role in the down-regulation of NF-kappa B activation, which is a key factor in the pathogenesis of IBD and is thus an excellent therapeutic target for the treatment of IBD.
Animals ; Carbon Monoxide ; Cobalt ; Colitis ; Colon* ; Cytokines ; Down-Regulation* ; Epithelial Cells ; Epithelium* ; Heme Oxygenase-1* ; Heme* ; HT29 Cells ; Humans ; Inflammation* ; Inflammatory Bowel Diseases ; Interleukin-1beta ; Iron ; Metabolism ; Mice ; NF-kappa B ; Tissue Donors ; Transcription Factors* ; Tumor Necrosis Factor-alpha

Human mesenchymal stem cells (hMSCs) are multipotent stem cells that can differentiate into several mesenchymal lineage cells. In this study, we established conditions that allowed a long term expansion of hMSCs. To search for the optimum culture condition, growth rates of hMSCs were measured in the presence of several growth factors. Hepatic growth factor (HGF) and leukemia inhibitory factor (LIF) did not facilitate proliferation of hMSCs. In contrast, basic fibroblast growth factor (bFGF) effectively promoted growth of the cells in vitro by 3 fold. The growth stimulatory effect of bFGF was dependent on the concentration. The adipogenic potential was dramatically decreased in hMSCs isolated from an aged donor whereas osteogenic potential was minimally decreased. Addition of bFGF resumed the adipogenic and osteogenic differentiation potential. Thus, the cells that expanded in the presence of bFGF retained the potential to differentiate into adipogenic, chondrogenic, or osteogenic lineage cells. MSCs could be expanded for at least 8 passages with bFGF and the resulting cells retained the normal karyotype. The cells were positive for CD9, CD13, CD15, CD90, CD137, and CD140b; but negative for CD14, CD34, and CD45. Importantly, the cells were found to express a neural stem cell marker, nestin, and a neuronal marker, beta-tubulin III. The results suggest that bFGF promote proliferation while maintaining multi-lineage differentiation potency of hMSCs. Finally, we suggest that it is critical to identify novel markers other than nestin or beta-tubulin III to monitor acquisition of neuronal phenotypes by hMSCs.
Fibroblast Growth Factor 2* ; Humans* ; Intercellular Signaling Peptides and Proteins ; Karyotype ; Leukemia Inhibitory Factor ; Mesenchymal Stromal Cells* ; Multipotent Stem Cells ; Nestin ; Neural Stem Cells ; Neurons ; Phenotype ; Tissue Donors ; Tubulin

Discovery of Nod2 has brought to light the significance of mononuclear cells as well as epithelial cells in inflammatory bowel disease (IBD) pathogenesis. Similarly, CCL20 is expressed in both mononuclear cells and epithelial cells and is likely to link innate and acquired immunity. We therefore asked whether CCL20 expression is altered in the peripheral blood mononuclear cells (PBMCs) from patients with ulcerative colitis (UC), a major type of IBD in Korea, and is correlated with the disease activity. The expression levels of CCL20 mRNA were significantly high in the PBMCs from the patients with UC. CCL20 protein expression was also up-regulated in the mucosal epithelium in UC but not in normal controls. Interestingly, however, disease activity index (DAI) revealed that untreated UC groups express higher expression levels of CCL20 mRNA than treated UC groups, implying that CCL20 may be a potential target for the anti-inflammatory treatments. In an agreement with this, three months follow up study revealed that the UC patients who were treated with 5-amino salicylic acid (5-ASA) and glucocorticoid showed dramatic decrease in their CCL20 mRNA levels as compared to untreated ones. Moreover, TNF-alpha-or IL-1beta-induced CCL20 secretion in human epithelial HT-29 cells was significantly diminished by the treatment with 5-ASA and/or dexamethasone, suggesting that CCL20 may be one of the central targets of the anti-inflammatory drugs. Collectively, these results suggest that CCL20 expression in UC may be associated with altered immune and inflammatory responses in the blood as well as the intestinal mucosa and further implied a potential for CCL20 as an important diagnostic marker for UC.
Adaptive Immunity ; Blood Cells* ; Colitis, Ulcerative* ; Crohn Disease ; Dexamethasone ; Epithelial Cells ; Epithelium ; Follow-Up Studies ; Gene Expression* ; HT29 Cells ; Humans ; Inflammatory Bowel Diseases ; Intestinal Mucosa ; Korea ; RNA, Messenger ; Salicylic Acid ; Sulfasalazine* ; Ulcer*

Various factors such as senescence, stress, neurodegenerative diseases including Alzheimer's disease (AD) contribute to the impairments of organs, especially brain. Also, they should be negative factors on normal brain function, like as memory and cognition. In this study, the neuroprotective role of BF-7, extracted from Bombyx mori, was examined agaist scopolamine-induced neurotoxicity in SK-N-SH cells. In order to know if the BF-7 has positive role on the cognition and memory, we examined using SD rat model and human. Scopolamine-induced memory impairments were observed, as measured by the passive avoidance and water maze tests, but treatment with BF-7 significantly improved memory and cognitive function. Moreover, the memory index and memory preservation of clinical experiments using MMSE-K tests were significantly improved memory and cognitive function. This results strongly represent that the BF-7 play effectively positive role in the improvement of brain function including learning and memory. Taken together, our results suggested that the BF-7 should be useful for developing strategies protecting nervous system and improving brain function.
Aging ; Alzheimer Disease ; Bombyx ; Brain ; Cognition ; Humans ; Learning ; Memory* ; Models, Animal ; Nervous System ; Neurodegenerative Diseases ; Scopolamine Hydrobromide